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The in-gel detection of proteins for various proteomic experiments is commonly done with the fluorescent Ru-II tris(bathophenanthroline disulfonate) complex (Ru(BPS)(3)), which is more cost-effective compared to commercial Ru-based formulations but requires tedious procedures for its preparation and strongly acidic staining conditions. Herein, we report the synthesis and characterization of heteroleptic Ru-II complexes Ru(BPS)(2)(BP) and Ru(BPS)(BP)(2) containing bathophenanthroline (BP) and bathophenanthroline disulfonate disodium salt (BPS) in comparison with Ru(BPS)(3). It was shown by fluorescent and UV-vis measurements that novel Ru-II complexes were excitable in both UV and visible light, close to emission bands of classical lasers, which is important for successful in-gel protein detection. Novel fluorescent dyes demonstrated improved protein detection in comparison with commercially available SYPRO Ruby staining solution. In addition, unlike commonly used staining protocols, staining with Ru(BPS)(BP)(2) can be performed at nearly neutral pH, thereby reducing artificial post-translational modifications (PTMs).

Cerium, an abundant lanthanide element, is widely used in human industry. The accumulation of Ce4+ ion, however, will damage the environment and biological organism. Therefore, its facile detection is highly needed. Herein, we design a hybrid sensing platform consisting of carbon dots (C-dots) and bathophenanthroline-disulfonate-Fe2+ complex (Bphen-Fe2+) for trace-level determination of Ce4+. Based on inner filter effect (IFE), the red-colored Bphen-Fe2+ complex severely quenches the fluorescence of C-dots. After addition of Ce4+, Fe2+ is oxidized to Fe3+, and the colorless Bphen-Fe3+ complex generates, which weakens the IFE efficiency and leads to the fluorescence recovery of C-dots. Meanwhile, due to the decreasing amount of Bphen-Fe2+ upon Ce4+ addition, the red color of the solution gradually fades, which enables visual detection of Ce4+ by the naked eyes. Under the optimized conditions, the C-dots/Bphen-Fe2+ system realizes the fluorometric and colorimetric sensing of Ce4+ in the range of 0.5–100 and 1.9–80 μM, with the limits of detection as low as 0.5 and 1.9 μM, respectively. This method also shows high selectivity over other common ions, and has an excellent applicability for monitoring of Ce4+ in real water samples.

13 Pags.- 1 Tabl.- 7 Figs., Main conclusion Fe uptake machinery of chloroplasts prefers to utilise Fe(III)–citrate over Fe–nicotianamine complexes., Iron uptake in chloroplasts is a process of prime importance. Although a few members of their iron transport machinery were identifed, the substrate preference of the system is still unknown. Intact chloroplasts of oilseed rape (Brassica napus) were purifed and subjected to iron uptake studies using natural and artifcial iron complexes. Fe–nicotianamine (NA) complexes were characterised by 5 K, 5 T Mössbauer spectrometry. Expression of components of the chloroplast Fe uptake machinery was also studied. Fe(III)–NA contained a minor paramagnetic Fe(II) component (ca. 9%), a paramagnetic Fe(III) component exhibiting dimeric or oligomeric structure (ca. 20%), and a Fe(III) complex, likely being a monomeric structure, which undergoes slow electronic relaxation at 5 K (ca. 61%). Fe(II)–NA contained more than one similar chemical Fe(II) environment with no sign of Fe(III) components. Chloroplasts preferred Fe(III)–citrate compared to Fe(III)–NA and Fe(II)–NA, but also to Fe(III)–EDTA and Fe(III)–o,o′EDDHA, and the Km value was lower for Fe(III)-citrate than for the Fe–NA complexes. Only the uptake of Fe(III)–citrate was light-dependent. Regarding the components of the chloroplast Fe uptake system, only genes of the reduction-based Fe uptake system showed high expression. Chloroplasts more efectively utilize Fe(III)–citrate, but hardly Fe–NA complexes in Fe uptake., This work was supported by the grants fnanced by the National Research, Development and Innovation Ofce, Hungary (NKFIH PD-112047, PD-111979 and K-124159), VEKOP2.3.3-15-2016-00008 and the Spanish Ministry of Economy and Competitiveness (MINECO; project AGL2016-75226-R, co-fnanced with FEDER). Á.S. was also supported by the Bolyai János Research Scholarship of the Hungarian Academy of Sciences (BO/00207/15/4). The authors (JP, LM, and RZ) gratefully acknowledge the fnancial support provided by the project LO1305 of the Ministry of Education, Youth and Sports of the Czech Republic.

A simple visual method for determining the total iron in human serum is proposed based on color development in the nanospace of mesoporous silica MCM-41 and a chromogenic ligand bathophenathroline disulfonate (BPS). Observing the color intensity of a complex between iron(II) and BPS devloped on the MCM-41 material by the naked eye enabled us to quntify iron(II) with a detection limit of 0.5 μM. The BPS-loaded MCM-41 was successfully applied for quantifying the total iron in human serum.

Background Free, non-protein bound, Fe(II), which can catalyse the formation of the toxic highly-reactive oxygen species (hROS), has been implicated in several neurodegenerative conditions. The determination of free Fe(II) and Fe(III) in samples obtained from microdialysis experiments has been limited by the small amounts of sample available. New method This work describes the development of a HPLC, with absorbance detection, method, based on the complexation of Fe(II) with bathophenanthroline disulfonate (BS), which allows a complete extracellular iron analysis with the small sample amounts that are available from in vivo microdialysis in rat brain. Results Microdialysis experiments using 6-hydroxydopamine stimulation, showed that basal-as well as evoked levels of extracellular Fe(II) and total iron could be determined in parallel with measurements of hROS formation. Comparison with existing methods Although a spectrophotometric BS-based assay has been reported for use in microdialysis samples from large animals, the present procedure is applicable to the small sample sizes available from studies in rat brain. It is simpler than the alternative, involving inductively-coupled plasma mass spectrometry. Conclusions The procedure described is simple and sensitive, giving a linear response in the Fe(II) concentration range of 50 −2000 nM. A 20 min microdialysis sample (flow-rate 3 μl/min) yields sufficient material for triplicate determinations of the evoked release of Fe(II) and total iron whilst leaving sufficient sample volume for determining hROS and amine or amino-acid neurotransmitter release.

Iron toxicity is a problem in many areas of wetland rice. Since Fe2+ is considered to be the toxic form of iron, the objective of this research was to determine the Fe2+ concentration in rice leaves using the chelator bathophenanthroline disulfonate (BPDS), disodium salt alone or combined with the chelator ethylenediaminetetraacetate (EDTA), disodium salt, where BPDS should solely chelate the Fe2+ and EDTA chelate only Fe3+. Thus, the combination of these chelators should stabilize the Fe oxidation states. It was also tested whether the chelators BPDS and EDTA could stabilize the oxidation states of Fe during the extraction of rice leaves. Extractions of rice leaves were carried out using an 1 mM BPDS or BPDS‐EDTA extractant solution. To test the stabilization of the Fe oxidation states by the combination of BPDS with EDTA, the extraction solution for one part of the samples contained 0.07 mM Fe3+. An extraction without plant material as control was also taken into consideration. The results indi...

When nonlethal levels of 2,4,5-trichlorophenol (0.2 mM) and the Cu(II)-bis(1,10-phenanthroline) complex [Cu(II)(OP)2] (0.1 microM) were combined, a remarkable synergistic cytotoxicity was observed as measured by the extent of bacterial inactivation. In contrast, no such synergism was observed for the combination of 2,4,5-trichlorophenol with the Cu(II)-bis(bathophenanthroline disulfonate) complex [Cu(II)(BPS)2] which has a chemical structure similar to Cu(II)(OP)2, except for the net charge. The synergism observed for 2,4,5-trichlorophenol and Cu(II)(OP)2 was found to be due to the neutralization of their opposite charge and formation of a lipophilic ternary complex which facilitated copper transport into the bacterial cells.

Direct determination of Fe(II) and Fe(III) is a fundamental technology for environmental Fe chemistry research. The key to accurate determination of Fe(II) and Fe(III) in soil, water, and plant samples is to stabilize Fe oxidation states during Fe extraction and analysis, but the goal has only been partially achieved in previous research. In this research, an approach to stabilizing the original oxidation states of Fe during extraction and analysis was studied. The following sequential procedures were conducted: computer simulation → colorimetric tests → Fe(III) reduction resistance tests → metal interference tests → sensitivity tests → determination of optimal pH for Fe extraction. Results indicated that EDTA (ethylenediamine-N,N,N′,N′-tetraacetate, disodium salt) and BPDS (bathophenanthroline disulfonate, disodium salt) were the most effective chelator pair with which to stabilize Fe oxidation states and that they may allow accurate, simultaneous determination of Fe(II) and Fe(III). This research was conducted at the Dep. of Soil, Crop and Atmospheric Sciences, Cornell Univ., Ithaca, NY.

International audience; Fluorescent detection of proteins is a popular method of detection allying sensitivity, linearity and compatibility with mass spectrometry. Among the numerous methods described in the literature, staining with ruthenium II tris(bathophenanthroline disulfonate) is particularly cost-effective, but slightly cumbersome owing to difficulties in the preparation of the complex and complexity of staining protocols. We describe here the modifications on both aspects that allow to perform a higher contrast staining and offer a more robust method of complex preparation, thereby maximizing the advantages of the method.

In order to clarify the mechanism of growth of musty-odor producing cyanobacteria, the effect of seven chelating agentssuch as EDTA (Ethylenediaminetetraacetic acid), CyDTA (Trans-1, 2-cyclohexanediamine-N, N, N′, N′-tetraacetic acid), DTPA (Diethylenetriamine-N, N, N′, N″, N″-pentaacetic acid), EDDHA {Ethylenediaminedi (o-hydroxyphenylacetic acid) }, HBED {N, N′-di (2-hydroxylbenzyl) -ethylenediamine-N, N′-diacetic acid}, DESF (Desferrioxamine B), BPDS (Bathophenanthroline disulfonate) on the growth ofPhormidium tenuewere studied using laboratory cultures.P.tenuegrew well in the pfesence of 1: 1 chelates (molar ratio of chelator to Fe) of CyDTA, DTPA, EDDHA and HBED as wellas EDTA as the iron sources.P. tenuegrew well in the presenceof CyDTA up to 17-fold for Fe (molar ratio) . However, the growth ofP.tenuewas suppressed in the presence of DTPA of 7-fold, EDDHA of 3-fold and HBED of 3-fold for Fe (molar ratio), respectively.P.tenuealso could utilize FeII (BPDS) 3chelate as the iron source. In addition, it could utilize FeII (BPDS) 3in the presence of HBED. However, it could not utilize FeII (BPDS) 3in the presence of DTPA, DESF or EDDHA.The present results indicate thatP.tenuecan utilize a wide variety of iron chelates as the iron sources and has a stronger ability of chelating functions on the cell surfaces compared withO.tenuisandA. macrospora. However, the addition of excess of chelating agents (DTPA, DESF, EDDHA and HBED) having higher stability constants of Fe (III) chelates affected the growth ofP.tenue. Furthermore, the results show that the absorption of iron byP.tenuedoes not only depend on the stability constants of Fe chelates and that it also depends on the chemical structure or characteristics of chelators.

In order to clarify the mechanism of growth of a musty-odor producing cyanobacterium Oscillatoria brevis isolated in Norway, the effects of iron forms on the growth and geosmin production of O.brevis were studied using laboratory cultures.O.brevis grew well and produced geosmin in the EDTA-free CT medium as well as in the presence of EDTA.Thus, O.brevis showed an ability to utilize colloidal iron and even iron oxides for its growth. This suggests that O.brevis would be expected to produce a specific high affinity iron chelator (siderophore) in the chelator-free medium.In the presence of bathophenanthroline disulfonate (BPDS), a strong Fe (II) chelating agent, O.brevis was able to grow.Furthermore, O.brevis was capable of growing in the medium containing a microbial siderophore, desferrioxamine B (DESF), instead ofEDTA.These results show that O.brevis has an excellent ability to utilize a variety of iron forms including Fe (III) -EDTA, Fe (II) -BPDS, Fe (III) -DESF chelates, colloidal ferric hydroxide and more thermodynamically stable crystalline iron oxides (Fe2O3, Fe3O4) in comparison with musty-odor producing cyanobacteria such as Anabaena macrospora, Phormidium tenue and Oscillatoria tenuis isolated from Lake Biwa

Pre-cast bis(2-hydroxyethyl)iminotris(hydroxymethyl)methane (Bis-Tris) gels have proven to be very suitable for pre-fractionation for LC-MS/MS analysis due to high reliability and long stability. To visualize proteins within gels fluorescence dyes proved to be a good tradeoff between sensitivity and MS-compatibility. The custom-made ruthenium dye represents a low-cost alternative regarding fluorescence-based protein visualization with high sensitivity. We demonstrate, that this dye is incompatible with Bis-Tris gels, while using Tris-Glycine gels a competitive sensitivity to commercially available stains can be achieved.

This study presents information on the dynamics of sugar beet (Beta vulgaris L.) root metabolic responses to Fe deficiency. Four days after Fe withdrawal from the nutrient solution, the concentration of reduced glutathione (GSH) and the activities of ferric chelate reductase (FC-R, E.C. 1.6.99.13) and ascorbate peroxidase (APX, E.C. 1.11.1.11) showed marked differences with the control values. The differences in root fresh weight, the concentrations of ascorbate (Asc) and soluble proteins, and proton release rates by roots became more prominent after the first week. Time course kinetics of FC-R activity and proton extrusion showed induction maxima, followed by rapid declines with the progress of Fe deficiency. The time courses of root Asc and GSH concentrations showed gradual increases with the advancement of Fe deficiency suggesting that plant responses to Fe deficiency stress may include a shift in the redox balance inside the root cells towards more reduced state. Data imply that an alternative Asc-dependent Fe reduction may become important with the advancement of Fe deficiency stress, when FC-R activity is declining. The time course of APX activity reveals that it could be a sensitive indicator of the initial changes in the plant Fe nutritional status., Work supported by grant BOS2001-2343 from the Spanish Ministry of Science and Technology to J.A. T.Z. was supported by a sabbatical grant (SAB95-0551) from the Spanish Ministry of Science and Education.

Water soluble palladium(II) complexes of bidentate diamine ligands, such as bathophenanthroline disulfonate, are stable, recyclable catalysts for the selective aerobic oxidation of terminal olefins to the corresponding alkan-2-ones in a biphasic liquid–liquid system.